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Volume 154, Issue 12

Capsule polysaccharide is a bacterial decoy for antimicrobial peptides Free

Abstract

Antimicrobial peptides (APs) are important host weapons against infections. Nearly all APs are cationic and their microbicidal action is initiated through interactions with the anionic bacterial surface. It is known that pathogens have developed countermeasures to resist these agents by reducing the negative charge of membranes, by active efflux and by proteolytic degradation. Here we uncover a new strategy of resistance based on the neutralization of the bactericidal activity of APs by anionic bacterial capsule polysaccharide (CPS). Purified CPSs from K2, serotype 3 and increased the resistance to polymyxin B of an unencapsulated mutant. Furthermore, these CPSs increased the MICs of polymyxin B and human neutrophil -defensin 1 (HNP-1) for unencapsulated , and PAO1. Polymyxin B or HNP-1 released CPS from capsulated , serotype 3 and overexpressing CPS. Moreover, this material also reduced the bactericidal activity of APs. We postulate that APs may trigger the release of CPS, which in turn will protect bacteria against APs. We found that anionic CPSs, but not cationic or uncharged ones, blocked the bactericidal activity of APs by binding them, thereby reducing the amount of peptides reaching the bacterial surface. Supporting this, polycations inhibited such interaction and the bactericidal activity was restored. We postulate that trapping of APs by anionic CPSs is an additional selective virulence trait of these molecules, which could be considered as bacterial decoys for APs.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): AP, antimicrobial peptide , CPS, capsule polysaccharide and HNP-1, human neutrophil α-defensin 1
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2008-12-01
2024-04-19
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References

  1. Appleyard R. K. 1954; Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from Escherichia coli K12. Genetics 39:440–452
     [Google Scholar]
  2. Arakawa Y., Wacharotayankun R., Nagatsuka T., Ito H., Kato N., Ohta M. 1995; Genomic organization of the Klebsiella pneumoniae cps region responsible for serotype K2 capsular polysaccharide synthesis in the virulent strain Chedid. J Bacteriol 177:1788–1796
     [Google Scholar]
  3. Arrecubieta C., Lopez R., Garcia E. 1994; Molecular characterization of cap3A, a gene from the operon required for the synthesis of the capsule of Streptococcus pneumoniae type 3: sequencing of mutations responsible for the unencapsulated phenotype and localization of the capsular cluster on the pneumococcal chromosome. J Bacteriol 176:6375–6383
     [Google Scholar]
  4. Beiter K., Wartha F., Hurwitz R., Normark S., Zychlinsky A., Henriques-Normark B. 2008; The capsule sensitizes Streptococcus pneumoniae to neutrophil alpha-defensins HNP 1–3. Infect Immun 76:3710–3716
     [Google Scholar]
  5. Bengoechea J. A., Lindner B., Seydel U., Díaz R., Moriyón I. 1998; Yersinia pseudotuberculosis and Yersinia pestis are more resistant to bactericidal cationic peptides than Yersinia enterocolitica . Microbiology 144:1509–1515
     [Google Scholar]
  6. Bengoechea J. A., Najdenski H., Skurnik M. 2004; Lipopolysaccharide O antigen status of Yersinia enterocolitica O: 8 is essential for virulence and absence of O antigen affects the expression of other Yersinia virulence factors. Mol Microbiol 52:451–469
     [Google Scholar]
  7. Bitter T., Muir H. M. 1962; A modified uronic acid carbazole reaction. Anal Biochem 4:330–334
     [Google Scholar]
  8. Boucher J. C., Yu H., Mudd M. H., Deretic V. 1997; Mucoid Pseudomonas aeruginosa in cystic fibrosis: characterization of muc mutations in clinical isolates and analysis of clearance in a mouse model of respiratory infection. Infect Immun 65:3838–3846
     [Google Scholar]
  9. Bragonzi A., Worlitzsch D., Pier G. B., Timpert P., Ulrich M., Hentzer M., Andersen J. B., Givskov M., Conese M., Doring G. 2005; Nonmucoid Pseudomonas aeruginosa expresses alginate in the lungs of patients with cystic fibrosis and in a mouse model. J Infect Dis 192:410–419
     [Google Scholar]
  10. Brogden K. A. 2005; Antimicrobial peptides: pore formers or metabolic inhibitors in bacteria?. Nat Rev Microbiol 3:238–250
     [Google Scholar]
  11. Campos M. A., Vargas M. A., Regueiro V., Llompart C. M., Alberti S., Bengoechea J. A. 2004; Capsule polysaccharide mediates bacterial resistance to antimicrobial peptides. Infect Immun 72:7107–7114
     [Google Scholar]
  12. Campos M. A., Morey P., Bengoechea J. A. 2006; Quinolones sensitize gram-negative bacteria to antimicrobial peptides. Antimicrob Agents Chemother 50:2361–2367
     [Google Scholar]
  13. Cobb L. M., Mychaleckyj J. C., Wozniak D. J., Lopez-Boado Y. S. 2004; Pseudomonas aeruginosa flagellin and alginate elicit very distinct gene expression patterns in airway epithelial cells: implications for cystic fibrosis disease. J Immunol 173:5659–5670
     [Google Scholar]
  14. Corsaro M. M., De C. C., Naldi T., Parrilli M., Tomas J. M., Regue M. 2005; 1H and 13C NMR characterization and secondary structure of the K2 polysaccharide of Klebsiella pneumoniae strain 52145. Carbohydr Res 340:2212–2217
     [Google Scholar]
  15. Díaz-Aparicio E., Aragón V., Marín C., Alonso B., Font M., Moreno E., Pérez-Ortiz S., Blasco J. M., Diaz R., Moriyón I. 1993; Comparative analysis of Brucella serotype A and M and Yersinia enterocolitica O : 9 polysaccharides for serological diagnosis of brucellosis in cattle, sheep, and goats. J Clin Microbiol 31:3136–3141
     [Google Scholar]
  16. Fresno S., Jimenez N., Izquierdo L., Merino S., Corsaro M. M., De C. C., Parrilli M., Naldi T., Regue M., Tomas J. M. 2006; The ionic interaction of Klebsiella pneumoniae K2 capsule and core lipopolysaccharide. Microbiology 152:1807–1818
     [Google Scholar]
  17. Frick I. M., Akesson P., Rasmussen M., Schmidtchen A., Bjorck L. 2003; SIC, a secreted protein of Streptococcus pyogenes that inactivates antibacterial peptides. J Biol Chem 278:16561–16566
     [Google Scholar]
  18. Ganz T. 1987; Extracellular release of antimicrobial defensins by human polymorphonuclear leukocytes. Infect Immun 55:568–571
     [Google Scholar]
  19. Groisman E. A. 1994; How bacteria resist killing by host-defence peptides. Trends Microbiol 2:444–449
     [Google Scholar]
  20. Guina T., Yi E. C., Wang H., Hackett M., Miller S. I. 2000; A PhoP-regulated outer membrane protease of Salmonella enterica serovar Typhimurium promotes resistance to α-helical antimicrobial peptides. J Bacteriol 182:4077–4086
     [Google Scholar]
  21. Gunn J. S., Miller S. I. 1996; Pho-PhoQ activates transcription of pmrAB, encoding a two-component regulatory system involved in Salmonella typhimurium antimicrobial peptide resistance. J Bacteriol 178:6857–6864
     [Google Scholar]
  22. Gunn J. S., Lim K. B., Krueger J., Kim K., Guo L., Hackett M., Miller S. I. 1998; PmrA-PmrB-regulated genes necessary for 4-aminoarabinose lipid A modification and polymyxin resistance. Mol Microbiol 27:1171–1182
     [Google Scholar]
  23. Gutsmann T., Hagge S. O., David A., Roes S., Bohling A., Hammer M. U., Seydel U. 2005; Lipid-mediated resistance of Gram-negative bacteria against various pore-forming antimicrobial peptides. J Endotoxin Res 11:167–173
     [Google Scholar]
  24. Hancock R. E. W., Chapple D. S. 1999; Peptide antibiotics. Antimicrob Agents Chemother 43:1317–1323
     [Google Scholar]
  25. Hardy G. G., Caimano M. J., Yother J. 2000; Capsule biosynthesis and basic metabolism in Streptococcus pneumoniae are linked through the cellular phosphoglucomutase. J Bacteriol 182:1854–1863
     [Google Scholar]
  26. Jin T., Bokarewa M., Foster T., Mitchell J., Higgins J., Tarkowski A. 2004; Staphylococcus aureus resists human defensins by production of staphylokinase, a novel bacterial evasion mechanism. J Immunol 172:1169–1176
     [Google Scholar]
  27. Kamerling J. P. 2000; Pneumococcal polysaccharides: a chemical view. In Streptococcus Pneumoniae: Molecular Biology and Mechanisms of Disease pp 81–114 Edited by Tomasz A. Larchmont, NY: Mary Ann Liebert;
     [Google Scholar]
  28. Lehrer R. I., Rosenman M., Harwig S. S., Jackson R., Eisenhauer P. 1991; Ultrasensitive assays for endogenous antimicrobial polypeptides. J Immunol Methods 137:167–173
     [Google Scholar]
  29. Link A. J., Phillips D., Church G. M. 1997; Methods for generating precise deletions and insertions in the genome of wild-type Escherichia coli: application to open reading frame characterization. J Bacteriol 179:6228–6237
     [Google Scholar]
  30. Ma S., Selvaraj U., Ohman D. E., Quarless R., Hassett D. J., Wozniak D. J. 1998; Phosphorylation-independent activity of the response regulators AlgB and AlgR in promoting alginate biosynthesis in mucoid Pseudomonas aeruginosa . J Bacteriol 180:956–968
     [Google Scholar]
  31. Mantovani A., Bonecchi R., Locati M. 2006; Tuning inflammation and immunity by chemokine sequestration: decoys and more. Nat Rev Immunol 6:907–918
     [Google Scholar]
  32. Martin D. W., Schurr M. J., Mudd M. H., Govan J. R., Holloway B. W., Deretic V. 1993; Mechanism of conversion to mucoidy in Pseudomonas aeruginosa infecting cystic fibrosis patients. Proc Natl Acad Sci U S A 90:8377–8381
     [Google Scholar]
  33. Moore R. A., Bates N. C., Hancock R. E. 1986; Interaction of polycationic antibiotics with Pseudomonas aeruginosa lipopolysaccharide and lipid A studied by using dansyl-polymyxin. Antimicrob Agents Chemother 29:496–500
     [Google Scholar]
  34. Moskowitz S. M., Ernst R. K., Miller S. I. 2004; PmrAB, a two-component regulatory system of Pseudomonas aeruginosa that modulates resistance to cationic antimicrobial peptides and addition of aminoarabinose to lipid A. J Bacteriol 186:575–579
     [Google Scholar]
  35. Nassif X., Fournier J. M., Arondel J., Sansonetti P. J. 1989; Mucoid phenotype of Klebsiella pneumoniae is a plasmid-encoded virulence factor. Infect Immun 57:546–552
     [Google Scholar]
  36. Newton B. A. 1955; A fluorescent derivative of polymyxin: its preparation and use in studying the site of action of the antibiotic. J Gen Microbiol 12:226–236
     [Google Scholar]
  37. Nicolas P., Mor A. 1995; Peptides as weapons against microorganisms in the chemical defence system of vertebrates. Annu Rev Microbiol 49:277–304
     [Google Scholar]
  38. Nizet V. 2006; Antimicrobial peptide resistance mechanisms of human bacterial pathogens. Curr Issues Mol Biol 8:11–26
     [Google Scholar]
  39. Peschel A. 2002; How do bacteria resist human antimicrobial peptides?. Trends Microbiol 10:179–186
     [Google Scholar]
  40. Poschet J. F., Boucher J. C., Firoved A. M., Deretic V. 2001; Conversion to mucoidy in Pseudomonas aeruginosa infecting cystic fibrosis patients. Methods Enzymol 336:65–76
     [Google Scholar]
  41. Rahn A., Whitfield C. 2003; Transcriptional organization and regulation of the Escherichia coli K30 group 1 capsule biosynthesis ( cps) gene cluster. Mol Microbiol 47:1045–1060
     [Google Scholar]
  42. Regue M., Izquierdo L., Fresno S., Jimenez N., Pique N., Corsaro M. M., Parrilli M., Naldi T., Merino S., Tomas J. M. 2005; The incorporation of glucosamine into enterobacterial core lipopolysaccharide: two enzymatic steps are required. J Biol Chem 280:36648–36656
     [Google Scholar]
  43. Selsted M. E., Ouellette A. J. 1995; Defensins in granules of phagocytic and non-phagocytic cells. Trends Cell Biol 5:114–119
     [Google Scholar]
  44. Shafer W. M., Qu X., Waring A. J., Lehrer R. I. 1998; Modulation of Neisseria gonorrhoeae susceptibility to vertebrate antibacterial peptides due to a member of the resistance/nodulation/division efflux pump family. Proc Natl Acad Sci U S A 95:1829–1833
     [Google Scholar]
  45. Sherbrock-Cox V., Russell N. J., Gacesa P. 1984; The purification and chemical characterisation of the alginate present in extracellular material produced by mucoid strains of Pseudomonas aeruginosa . Carbohydr Res 135:147–154
     [Google Scholar]
  46. Spinosa M. R., Progida C., Tala A., Cogli L., Alifano P., Bucci C. 2007; The Neisseria meningitidis capsule is important for intracellular survival in human cells. Infect Immun 75:3594–3603
     [Google Scholar]
  47. Steinmoen H., Knutsen E., Havarstein L. S. 2002; Induction of natural competence in Streptococcus pneumoniae triggers lysis and DNA release from a subfraction of the cell population. Proc Natl Acad Sci U S A 99:7681–7686
     [Google Scholar]
  48. Steinmoen H., Teigen A., Havarstein L. S. 2003; Competence-induced cells of Streptococcus pneumoniae lyse competence-deficient cells of the same strain during cocultivation. J Bacteriol 185:7176–7183
     [Google Scholar]
  49. Straus D. C., Atkisson D. L., Garner C. W. 1985; Importance of a lipopolysaccharide-containing extracellular toxic complex in infections produced by Klebsiella pneumoniae . Infect Immun 50:787–795
     [Google Scholar]
  50. Takemura H., Kaku M., Kohno S., Hirakata Y., Tanaka H., Yoshida R., Tomono K., Koga H., Wada A. other authors 1996; Evaluation of susceptibility of gram-positive and -negative bacteria to human defensins by using radial diffusion assay. Antimicrob Agents Chemother 40:2280–2284
     [Google Scholar]
  51. Vaara M. 1992; Agents that increase the permeability of the outer membrane. Microbiol Rev 56:395–411
     [Google Scholar]
  52. Ventura C. L., Cartee R. T., Forsee W. T., Yother J. 2006; Control of capsular polysaccharide chain length by UDP-sugar substrate concentrations in Streptococcus pneumoniae . Mol Microbiol 61:723–733
     [Google Scholar]
  53. Westphal O., Jann K. 1963; Bacterial lipopolysaccharides extraction with phenol-water and further applications of the procedure. Methods Carbohydr Chem 5:83–91
     [Google Scholar]
  54. Wiese A., Gutsmann T., Seydel U. 2003; Towards antibacterial strategies: studies on the mechanisms of interaction between antibacterial peptides and model membranes. J Endotoxin Res 9:67–84
     [Google Scholar]
  55. Zilbauer M., Dorrell N., Boughan P. K., Harris A., Wren B. W., Klein N. J., Bajaj-Elliott M. 2005; Intestinal innate immunity to Campylobacter jejuni results in induction of bactericidal human beta-defensins 2 and 3. Infect Immun 73:7281–7289
     [Google Scholar]
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Capsule polysaccharide is a bacterial decoy for antimicrobial peptides
Microbiology 154, 3877 (2008); https://doi.org/10.1099/mic.0.2008/022301-0
/content/journal/micro/10.1099/mic.0.2008/022301-0
/content/journal/micro/10.1099/mic.0.2008/022301-0
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